The hemocytes collected from oysters were resuspended in L15 cell culture media (extra addition of 20.2 g/L NaCl, 0.54 g/L KCl, 0.6 g/L CaCl, 3.9 g/L MgCl2, and 1 g/L MgSO4) and deposited on dishes pre-coated with poly-L-lysine, and then incubated at 37°C for 1 h to adhere to the glass slides. The hemocytes on the object slides were fixed with 4% paraformaldehyde, washed with PBS, and permeabilized with 1% Triton X-100 for 5 min. After three times of PBS wash, the hemocytes were blocked with 3% (w/v) fetal bovine serum albumin (BSA) at 37°C for 30 min and incubated with the antibody [anti-CgIRF-8, anti-CgcGAS, 1:500 (v/v) in 3% BSA] at 37°C for 1 h. Then the samples were washed with PBS and incubated with Alexa Fluer 488-labeled goat-anti-mouse antibody (Solarbio life sciences, China) diluted at 1:1,000 (v/v) with 3% BSA at 37°C in the dark for 1 h. The DAPI dihydrochloride (1 mg/ml in PBS; Solarbio Life Sciences, China) was added to stain the nucleus for 5 min followed by three times wash with PBS. Fluorescence was observed using Carl Zeiss Axio Imager A2 microscope (Carl Zeiss, Germany).
Carl axio imager a2 microscope
Manufactured by Zeiss
Sourced in Germany
The Carl Zeiss Axio Imager A2 is a high-performance microscope designed for a variety of applications. It features a modular optical system, allowing for the integration of various imaging techniques such as brightfield, darkfield, and fluorescence. The microscope is equipped with a motorized stage and focus drive, providing precise control and positioning of the sample.
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Lab products found in correlation
4 protocols using carl axio imager a2 microscope
1
Immunofluorescence Analysis of Oyster Hemocytes
2
Suppressing Conidial Production and Germination in Cryptococcus destructans
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To suppress conidial production by bacterial treatment, C. destructans was cultured in V8 liquid medium with 1%, 0.1%, or 0.01% bacterial cultures at an optical density (OD) of 1.0 with shaking at 150 rpm, at 21℃, for 9 days. After inoculation for 3, 6, or 9 days, the number of spores was calculated using a hemocytometer. For the conidial germination assay, each bacterial culture was diluted serially (OD 1.0, 0.1, and 0.01), co-mixed with the conidium suspension (1 × 104 conidia/mL), placed on a hydrophobic slide (Knittel Glaser, Braunschweig, Germany), and incubated in a humidity box at 25℃. After incubation for 0.5, 1, 1.5, or 2 hr, the number of germinating spores was counted among 100 spores per treatment. To examine the inhibition of chlamydospore formation, we used Czapek-Dox broth treated with bacteria at an OD of 0.001. A mycelial agar plug (5 mm diameter) was inoculated into each medium, and the medium was incubated at 21℃ with shaking at 150 rpm. Chlamydospore production was observed after 5 days under a Carl Zeiss Axio Imager A2 microscope (Carl Zeiss, Oberkochen, Germany). Each experiment was replicated three times, and three separate experiments were conducted.
3
Microscopic Identification of Algae Genera and Species
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Microscopic morphologic identification at the genus level was performed according to Bellinger & Sigee, 2015 [56 ]. Further identification to species levels was accomplished by comparison with the species original descriptions that are available at the AlgaeBase [57 ]. In the case of the as of yet undescribed species, the morphological comparisons were made with the closest strains obtained in the molecular identification step: Desmodesmus sp. MAT2008c [58 (link)]; Micractinium sp. CCAP 211/92 [39 (link)]; Desmodesmus sp. GM4a [59 (link)]. A Carl Zeiss Axio Imager A2 microscope (Zeiss.co, Brazil) equipped with Differential Interference Contrast (DIC) was used for morphological analysis.
4
Histological Assessment of Cartilage Degeneration
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Histological changes were assessed to determine the effects of FP-MD and Celecoxib on joint cartilage degeneration. After 3 weeks post-OA induction, the rats were euthanized with CO2, and the affected knee joints were dissected and fixed in 10% formalin for 24 h at 4 °C, decalcified with 5% formic acid for 2 weeks, and then dehydrated in graded acetone and embedded in paraffin. Sections (thickness, 4–5 μm) were stained with hematoxylin and eosin (H&E) and Safranin-O/Fast green to evaluate structural cartilage damage and the proteoglycan loss, respectively, and then observed under Carl Zeiss Axio Imager A2 microscope (Carl Zeiss, Deisenhofen, Germany). All stained slides were histologically evaluated and statistically graded on a scale of 0–13 by double-blind observation, according to the modified Mankin scoring system [37 (link)].
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